Saturation Mutagenesis Genome Engineering of Infective ΦX174 Bacteriophage via Unamplified Oligo Pools and Golden Gate Assembly

Here we present a novel protocol for the construction of saturation single-site—and massive multisite—mutant libraries of a bacteriophage. We segmented the ΦX174 genome into 14 nontoxic and nonreplicative fragments compatible with Golden Gate assembly. We next used nicking mutagenesis with oligonucl...

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Publicat a:ACS Synth Biol
Autors principals: Faber, Matthew S., Van Leuven, James T., Ederer, Martina M., Sapozhnikov, Yesol, Wilson, Zoë L., Wichman, Holly A., Whitehead, Timothy A., Miller, Craig R.
Format: Artigo
Idioma:Inglês
Publicat: 2020
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Article
Accés en línia:https://ncbi.nlm.nih.gov/pmc/articles/PMC7055157/
https://ncbi.nlm.nih.gov/pubmed/31825605
https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1021/acssynbio.9b00411
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